Uterine inflammation status modulates eggshell mineralization via calcium transport and matrix protein synthesis in laying hens

This study explored the effects of uterine inflammation on eggshell mineralization,ultrastructure and mechanical properties in laying hens modified by a lipopolysaccharide(LPS)challenge or dietary essential oil(EO)addition.In trial 1,a total of 72 Hy-line Brown layers at 36 wk of age were randomly assigned to 3 treatment groups(n=8),where they were intravenously injected with phosphate buffered saline,LPS at 1 mg/kg body weight,or LPS 3 times at 24-h intervals.In trial 2,a total of 288 Hy-line Brown layers at 60 wk of age were randomly divided into 4 groups(n=8),where they were fed basal diets supplemented with EO at 0,50,100 and 200 mg/kg for 12 wk.A uterine inflammation model was constructed with LPS treatment,indicated by the elevated expression of IL-1β and TNF-α(P<0.05)and lymphocyte infiltration.Uterine inflammation caused remarkable decreases in eggshell thickness and mechanical properties with structure deteriorations(P<0.05).Uterine inflammation stimulated the expression of matrix proteins ovotransferrin(TF)and ovalbumin(OVAL),while depressing the mRNA levels of calbindin-1(CALB1)and osteopontin in uterine mucosa(P<0.05).In contrast,EO addition alleviated uterine inflammation,evidenced by depressed levels of IL-1b and IL-6(P<0.05).There was a significant elevation in shell thickness and breaking strength following EO intervention(P<0.05),and these effects were maximized at addition of 100 mg/kg.Further,EO improved shell ultrastructure including more early fusion,less type B mammillae,and increased effective thickness(P<0.05).The alleviated inflammation decreased the expression of OVAL and TF,whereas ion transport genes like CALB1 and solute carrier family 26 member 9 were upregulated(P<0.05).Our findings suggest that inflam-matory status can impact uterine functions in calcium transport and the synthesis of matrix proteins especially such as OVAL and TF,which in turn modulates calcium precipitation and ultrastructure for-mation,thereby determining eggshell mechanical properties.These findings provide a novel insight into the uterine inflammation-mediated modifications of eggshell quality.